GSM6505188: H3K27me3, wt, E12.5, Pinna; Mus musculus; ChIP-Seq
Embryonic tissues were micro-dissected at E10.5, E12.5 or E14.5. Dissected tissue from E10.5 and E12.5 embryos was kept in 1× PBS on ice, then treated with 0.5% trypsin/1× EDTA at 37 °C for 10 minutes and immediately put on ice. Dissected tissue from E14.5 embryos was kept in 1× PBS on ice, then treated with papain digestion mix (10 mg ml−1 papain, 2.5 mM cysteine, 10 mM HEPES (pH 7.4), 0.5 mM EDTA and 0.9× DMEM) for 7 min at 37 °C and immediately put on ice. The tissue was rinsed once in ice-cold 1× DMEM/10% FBS, followed by two washes in ice-cold 1× DMEM. Tissue was dissociated by pipetting. Then the dissociated cells were cross-linked with 1% formaldehyde for 10 min at room temperature (RT) and quenched with 125 mM glycine for 5 min at RT. Cells were spun down (1600 rpm, 10 minutes, 4 °C). Cranial NCCs with genotype Wnt1::Cre;ROSARFP were filtered and collected by FACS. NEBNextUltra for Illumina